Preparation for use of aspartate and vitamin b12 or biotin for regulating ketone bodies

ABSTRACT

It has been found that high amounts of aspartate equivalents in combination with vitamin B12 and/or biotin, especially in relative absence of glutamate equivalents, improve the metabolism of ketobodies and/or lactate in a mammal&#39;s body, especially in diseased or traumatic conditions. As a result, levels of ketobodies and lactate can be decreased and unphysicologically high acidity normalised. Thus, it is an object of the invention to provide an enteral nutritional or a pharmaceutical composition for the treatment and/or prevention of disturbed ketone and lactate metabolism, i.e. elevated concentrations of ketone bodies, lactate and/or other organic acids and/or insufficient pH homeostasis, especially elevated concentrations of ketone bodies and/or lactate, in a mammal&#39;s blood, wherein the composition comprises high amounts of aspartate equivalents in combination with vitamin B12 and/or biotin, preferably in relative absence of glutamate equivalents.

RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.13/195,831, filed Aug. 1, 2011, which is a continuation of U.S. patentapplication Ser. No. 11/572,197, filed Mar. 15, 2007, now U.S. Pat. No.8,003,600, which is a National Stage Application under 35 U.S.C. §371 ofPCT/NL2005/000520, filed Jul. 18, 2005, which claims priority from U.S.Provisional Patent Application No. 60/588,793, filed Jul. 19, 2004,European Patent Application No. 04077100.8, filed Jul. 19, 2004 andEuropean Patent Application No. 04077359.0 filed Aug. 20, 2004. Theforegoing applications are incorporated herein by reference in theirentirety.

FIELD OF THE INVENTION

The invention pertains to a preparation for use of specific proteinand/or peptide fractions having a high aspartate content in combinationwith vitamin B12 and/or biotin for the prevention or treatment ofmetabolic disorders associated with elevated concentrations of ketonebodies, lactate and/or other organic acids, especially ketone bodies andlactate, in a mammal's blood.

BACKGROUND OF THE INVENTION

Under healthy conditions, carbohydrates are converted after consumptionto glucose, which is the body's primary source of energy. However, whenthe intake of carbohydrates is limited for a long enough period of time,or when the carbohydrate metabolism is disturbed, a point is reachedwhere the body draws on alternative energy systems, fat or amino acidstores, for fuel. Upon catabolism of the lipids, several metabolites maybe produced, like acetoacetate, acetone and β-hydroxybutyric acid, whichmetabolites are referred to as ketone bodies. These compounds serve asimportant metabolic fuels for many peripheral tissues, particularlyheart and skeletal muscle, and in the absence of glucose, ketone bodiesbecome the brain's major fuel sources.

When the formation of ketone bodies exceeds the mammal's capacity oftreating them, the ketone bodies are accumulated in blood to causeketonaemia. Conditions where the concentration of ketone bodies is highin urine are called ketonuria, and both of them are generally calledketosis. During ketosis or hyperketonaemia, ketone body levels in theblood become abnormally high. Severe ketosis may result in acidosis, acondition in which blood pH typically decreases below 7.3, the partialpressure of carbon dioxide (PCO₂) in blood is below 30 mm Hg andbicarbonate levels in blood below 15 mm Hg. The symptoms of acidosisinclude malaise, weakness, anorexia and vomiting and these mayeventually lead to coma and even death.

During a disturbed carbohydrate metabolism, as may occur during insulinresistance or during anaerobic conditions, pyruvate often is notsufficiently metabolised to Krebs cycle intermediates, but at leastpartially to lactic acid instead. Accumulation of the latter may occuron a local level, e.g. in tissue or muscle, wherein it will cause ametabolic disturbance of cell functioning and pain, or it may occursystemically which may lead to acidosis. Disturbed carbohydratemetabolism which leads to lactic acidosis can be associated with livermalfunction, either through liver damage or underdeveloped enzymaticfunctions therein, like may occur in part of the babies of younggestational age. In lactic acidosis, a condition in which lactate levelsin the blood are typically above 2 mmol/l. It is common to make adistinction between hyperlactacidemia and severe lactic acidosis interms of the lactate concentration in the blood, usually at about 5mol/l.

In addition to lactic acidosis or ketoacidosis there is a group oforganic acidurias (some 25-30 different types) that belongs to the groupof metabolic acidoses, wherein an organic acid accumulates in the bloodand urine.

Ketosis can occur due to high endogenous biosynthesis and/or impartedclearance or metabolism. Many people suffering from severe energymalnutrition or protein-energy malnutrition also experience a form ofketosis or from an even more severe form thereof, called ketoacidosis(ketone body blood levels above 7 mmol/l). Also diabetics sufferfrequently from abnormal high levels of ketone bodies. Ketosis or evenketoacidosis can be caused by inborn or temporary metabolism errors,e.g. errors in branched chain metabolism, like in maple syrup disease,during inherited disorders in glycogen synthesis rate, or in personshaving certain types of inherited errors in metabolism, e.g. personssuffering from propionic acidemia, isovaleric acidemia, methylmalonicacidemia, oxoacid coenzym A thiolase deficiency or deficiencies in theactivity of other thiolases, and persons having an underdevelopedmetabolic system like infants of young gestational age.

Ketosis can also play a role in persons suffering from hyperglycaemia,which is a metabolic state of the body wherein glucose levels in bloodare increased compared to normal concentrations. Yet, despite these highblood glucose levels, cells “starve” since the insulin-stimulatedglucose entry into cells is somehow impaired. Examples of personssuffering from hyperglycaemia are those which are diagnosed to sufferfrom the so-called metabolic syndrome or syndrome X, obesity and severaltypes of diabetes, like type I, type II and gestational diabetes.Especially those persons suffering from an imparted insulin-release orfrom “insulin resistance”, will frequently develop a ketosis state.

Long term hyperglycaemia resulting from disturbed carbohydratemetabolism will result in increased formation of undesired advancedglycation products (AGE) by interaction between reactive amino groups inproteins, like primary amino groups such as those occurring in lysine,and in particular aldehydes, e.g. those resulting from reducing sugars.In this way carboxymethylated lysines are formed. Maillard-typereactions of endogenous proteins like enzymes and structural proteinsimpart their function which results in undesired loss of function of thetotal organ or tissue. These complications are in particularcardiovascular problems, like macro- and micro-angiopathy, problems withthe liver, pancreas, kidney, skin, the eye but also embryopathy duringpregnancy are often observed in diabetics and some of them also in theaged population.

Where ketosis is caused by an impaired glucose metabolism, it is oftentreated by administering insulin or a sugar, such as glucose, xylitol,or the like. However, as discussed above, ketosis is not alwaysassociated with a disordered glucose metabolism or AGE formulation, andwhere a relation exists, the effect of these sugars and insulin on thereduction of the concentration of ketone bodies is often transitory andlasts only a short period of time.

Hyperlactacidemia can be due to a disorder in cellular respiration,abnormalities in the activity of pyruvate dehydrogenase, the Krebscycle, the respiratory chain or due to liver function problems includingdisorders in glycogen metabolism, gluconeogenesis and fatty acidoxidation. Lactic acidemias can also be observed during chronicinfections, in particular of the urinary tract, chronic diarrheas andtissue hypoxias as may occur during ischaemic events like those that areapplied during surgery or traumatic experiences where blood supply isinterrupted, but also in underdeveloped metabolic or anatomical systemslike in part of the neonates. During hyperlactacidemia the weight ratioof lactate to pyruvate in blood preprandially will typically be above0.35:1. It can be observed in a variety of acquired circumstancesincluding infections, severe catabolism, organ dysfunction and tissueischaemia, but also during some inherited metabolic disorders. It is acommon disease symptom in pediatrics. Ketonuria, hyperlactacidemia andhyperammonemia, but also abnormal values of pyruvate, glucose, bloodgases, electrolytes and pH are important indicators of the metabolicsituation of the patient.

A need exists for a nutritional preparation, supplement or a dieteticregimen for the prevention or treatment of metabolic disordersassociated with elevated concentrations of ketone bodies and/or lactatein blood, and/or advanced glycation products and/or Maillard products intissue in a mammal. Preferably consumption of the nutritionalpreparation or supplement should be easy to comply with, because of itsenjoyable organoleptic properties, it should fit in normal day life andfeeding/drinking practices and should have no undesired side effects.

Neonates and in particular preterm babies often suffer fromunderdeveloped metabolic systems, which need to adapt in a short time toa new nutritional regimen. During the first few days and even weeksdramatic changes occur in their body for example with regard toexpression of enzymes, organ capacity e.g. of the liver, pancreas, gutand kidneys and gut content. When nutritional practices are not adaptedto their metabolic capabilities, disorders and diseases can be observedsuch as abnormally high or low levels of lactate, ketone bodies, ammoniaand pH in the blood, which often require medical intervention.

Ketones may also be formed when high amounts of lipids or excessbranched chain amino acids are consumed. In particular when consumingcomplete nutritional formulae in which lipids provide more than 40energy percent for adults, or even more than 52 en % for prematureinfants, ketones can be formed. The same problem occurs when in completenutritional formulae the amount of branched chain amino acids is high,e.g. more than 24 g/100 g amino acids, or even more than 26 g. Completenutritional products for adult people provide per daily dose more than80 g protein and more than 1800 kcal. For premature infants completenutrition provides per daily dose 6 g protein and 225 kcal energy.

An inverse correlation between blood total ketone body and alanineconcentrations has been reported in the art. Nosadini, R. et al.published in Biochem J (1980), 190, 323-332 a model study in ratswherein it is demonstrated that after consumption of high doses ofalanine blood levels of ketone bodies were decreased. Its positiveeffect was attributed partly to the enhanced oxaloacetate availability,which in turn was thought to result in enhanced citrate formation anddecreased intramitochondrial acetyl-CoA availability for ketogenesis.

This effect was partially mimicked by using extremely high doses of 6mmol (=0.69 g) of aspartate per kg body weight per hour to 48-hoursstarved rats, whereas half a dosis did not yield any significant effecton blood ketone body concentrations. It is clear that in humans weighing70 kg equivalent doses of about 193 g aspartate per day would impartconsumption of the normal diet heavily and put high demands onwillingness to comply by the patient.

SUMMARY OF THE INVENTION

It has been found that high amounts of aspartate equivalents incombination with vitamin B12 and/or biotin, more preferably aspartateequivalents in combination with vitamin B12 and biotin, especially inrelative absence of glutamate equivalents, improve the metabolism ofketobodies and/or lactate in a mammal's body, especially in diseased ortraumatic conditions. As a result, levels of ketobodies and lactate canbe decreased and unphysicologically high acidity normalised.

Thus, it is an object of the invention to provide an enteral nutritionalor a pharmaceutical composition for the treatment and/or prevention ofdisturbed ketone and lactate metabolism, i.e. elevated concentrations ofketone bodies, lactate and/or other organic acids and/or insufficient pHhomeostasis, especially elevated concentrations of ketone bodies and/orlactate, in a mammal's blood, in particular in diseased, traumatized ormetabolically stressed state of a mammal, wherein the compositioncomprises high amounts of aspartate equivalents in combination withvitamin B12 and/or biotin, preferably in relative absence of glutamateequivalents.

With “elevated concentrations of ketone bodies and/or lactate” and“disturbed ketone and lactate metabolism” are understood concentrationsof ketone bodies comprising acetoacetate, acetone and β-hydroxybutyricacid higher than 0.5 mmol/l in blood, and concentrations of lactatehigher than 2 mmol/l in blood.

The composition preferably contains a protein fraction comprising atleast 10.8 wt % aspartate equivalents, based on the total weight of theprotein fraction.

It is preferred that at least part of the aspartate equivalents isprovided by an aspartate source containing at least 12.0 wt %,preferably at least 12.3 wt % aspartate equivalents. Preferably, theprotein fraction further comprises glutamate equivalents in a weightratio of aspartate equivalents to glutamate equivalents (asp:glu)between 0.41:1 and 5:1.

Such an aspartate source containing at least 12.0 wt % can be an intactprotein, a protein isolate, concentrate or hydrolysate, and/or freeaspartate equivalents. If the aspartate source containing at least 12.0wt % is a protein, a protein isolate, concentrate of hydrolysate, it ispreferred that it is present in an amount of 5-100 wt %, more preferably8-70 wt %, even more preferably 10-60 wt % of the protein fraction. Whenmore than one protein containing at least 12.0 wt % aspartate ispresent, the above numbers apply to the sum of these proteins. In casethe aspartate source is formed from free aspartate equivalents, theseare preferably present in an amount of 0.2-9 wt %, more preferably 0.5-6wt %.

It is also an object of the invention to provide an enteral compositioncontaining 15-22 en % of a protein fraction and 25-50 en % of acarbohydrate fraction for the aforementioned use, wherein the proteinfraction comprises 10.8-30 wt % of aspartate equivalents, based on thetotal weight of the protein fraction, and wherein the compositionfurther contains at least one of vitamin B12 and biotin.

It is a further object of the invention to provide a method forpreventing or treating elevated blood concentrations of ketone bodies,lactate and/or other organic acids and/or insufficient pH homeostasis,especially elevated concentrations of ketone bodies and/or lactate, in amammal in need thereof, the method comprising administering the enteralnutritional or pharmaceutical composition of the invention to saidmammal.

DETAILED DESCRIPTION OF THE INVENTION Aspartate, Glutamate

The amino acids aspartic acid, asparagine and glutamic acid andglutamine are considered as non-essential amino acids in mammals, as themammal body has a metabolic capacity to synthesise these amino acidswhen necessary. The physiological active isomer is the L-form and theendogenous amino acids are typically in equilibrium with theirketo-analogues oxaloacetate (for aspartate) and alpha-ketoglutarate (forglutamate). Asparagine and glutamine can be hydrolysed in the body torespectively aspartate and glutamate under release of an ammonia groupvia interaction with the enzymes asparaginase and glutaminase,respectively. Aspartate and glutamate are sometimes considered asneurotoxins.

During the application of standard analysis methods for amino acidcontents, asparagine and glutamine are easily hydrolysed, which is whyoften in amino acid compositions of proteins no separate amount ofasparagine is provided but instead an amount for the sum of asparagineand aspartate is given. The same applies to glutamine.

For the purpose of this document, “aspartate equivalents” are defined ascomponents that are able to release L-aspartate in the body, eitherdirectly or after digestion, absorption, and metabolic conversion by theliver in situations in which the equivalent has been consumed orally orenterally, for example via tube feeding. Examples of aspartateequivalents are proteins or peptides that comprise L-aspartic acidand/or L-asparagine, free amino acids, either synthesised or extractedfrom natural materials, salt forms of the free amino acids, for examplesalts with metal ions like sodium, potassium, zinc, calcium, magnesiumor with other compounds like other amino acids, carnitine, taurine, orquaternary ammonium compounds like choline or betaine, the esterifiedforms of the amino acids, like those compounds which comprise an acylmoiety bound to one of the carboxylic acid moieties, or esters resultingfrom organic molecules like pyruvic acid, and derivatives of the freeamino acids in which an alkyl or acyl group has been attached to theprimary nitrogen atom. Thus, aspartate equivalents comprise any compoundhaving the formula R¹—NH—CH(COR²)—[CH₂]_(n)—CO—OR³ orR¹—NH—CH(COR²)—[CH₂]_(n)—CO—NHR³, wherein n=1, R¹ is H, (substituted)alkyl, or acyl (including C-peptidyl), R² is OH, OR³, NHR³ orN-peptidyl, and R³ is H, (substituted) alkyl or acyl, as well as theanion and cation salts and zwitterions. The same applies to glutamateequivalents, with the exception that n=2. The peptides are preferablyobtained by hydrolysis of intact protein. The keto analoguesoxaloacetate and its derivatives are less suitable forms for inclusionin a nutritional product due to technological (processing) and stabilityproblems that may arise.

Dosages are given in grams of L-aspartic acid. Equivalent dosages ofalternative components can be calculated by using the same molar amountand correcting for the molecular weight of the alternative component. Inthe calculations the residues in peptides and proteins are corrected forthe lack of a water molecule in the amino acid chain. All equivalentscontribute to the total weight in their complete, i.e hydrolysed form,including the water molecule.

“Glutamate equivalents” are defined in a similar way as for theaspartate equivalents. They include proteins or peptides that compriseL-glutamic acid and/or L-glutamine, free glutamate and glutamine aminoacids, either synthesised or extracted from natural materials, saltforms of the free amino acids etc. N-Acetyl glutamine and N-acetylglutamate are also suitable forms. Throughout the description and claimsdosages are given in grams of L-glutamine, the equivalents corrected forthe lacking water molecule in the case of peptide and proteinconstituents.

The amounts of aspartate and glutamate equivalents are calculated on thebasis of the total nutritional or pharmaceutical composition. In casethe composition consists of different portions, the amounts of thoseequivalents in the different portions are to be added.

Throughout this document, “free aspartate equivalents” or “freeglutamate equivalents” are understood to comprise aspartate, asparagine,glutamate and glutamine, and their free acid as well as their anionicforms and salts, such as alkali metal salts, alkaline earth metal salts,ammonium salts, substituted ammonium salts and zwitterionic species; theacids are indifferently referred to by their acid name or their anionicnames, e.g. aspartic acid or aspartate and glutamic acid or glutamate,respectively. Free aspartate equivalents and free glutamate equivalentsalso include dipeptides containing at least one molecule of aspartateand glutamate, respectively. The dipeptides serve as a source ofaspartate and glutamate, and should not have an independent biologicalaction in the concentration range wherein they are used.

However, it is preferred to use L-aspartic acid or its derivativesinstead of L-asparagine or its derivatives in order to preventproduction of undesired by-products during processing, in particularwhen the aspartate equivalents are included in free form, i.e. not asoligo- or polypeptide. Suitable forms of L-aspartic acid are salts withmetals like sodium, potassium, calcium, zinc and magnesium or with aminoacids like L-lysine and L-histidine.

The amount of aspartate salts is not allowed to exceed 9 wt %,preferably less than 6 wt % of the nutritional preparation, and inparticular the amount of each individual aspartate salt should notexceed 4.8 wt % in the case the protein fraction is administered to apatient in a liquid form, in order to avoid electrolyte imbalances. Forexample the amount of potassium will typically be less than 400,preferably 50-250 and most preferably 100-180 mg per 100 ml. The amountof magnesium will typically be less than 200 mg, preferably 10-120 andmore preferably 12-80 mg per 100 ml. Also dipeptides comprising one ormore aspartic acid moieties are suitable, though not preferredembodiments. Again an alternative source is plant extracts like extractsfrom sugar cane, especially those that are rich in aspartate and betaineor extracts from potato. By at least partial hydrolysis the aspartatefraction becomes more rapidly available to the patient.

Because of its importance in combating disorders related to elevatedconcentrations of ketone bodies, lactate and organic acids in blood, theamount of aspartate equivalents can be further increased, but theprotein fraction should contain no more than 95 wt %. The proteinfraction preferably comprises at least 10.8 wt % of aspartate,preferably 11.0-70 wt %, more preferably 11.5-50 wt %, even morepreferably 11.8-45 wt %, even more preferably 12.0-40 wt % and mostpreferably 12.5-36 wt %, in particular 12.8-30 wt % more in particularless than 25 wt % of aspartate equivalents, based on the weight of theprotein fraction. A protein fraction containing more than 13.0 wt %, oreven more than 14.0 wt % of aspartate equivalents is particularlypreferred.

A protein fraction according to the invention preferably comprises anaspartate fraction that is rapidly digested and absorbed and thereforeavailable in the body. This can be achieved by including at least partof the aspartate equivalents in a form that passes rapidly the stomach,and does not put high demands on the activity of digestive enzymes likepepsin, trypsin and chymotrypsin. In one embodiment it is thus preferredthat at least a part, preferably at least 0.2 wt %, more preferably atleast 0.5 wt %, even more preferably at least 0.7 wt % and mostpreferably at least 1.0 wt %, in particular at least 1.5 wt % of theaspartate equivalents are free aspartate equivalents and/or di-peptidescontaining at least one molecule of aspartate equivalents.

Further, the protein fraction of the invention preferably comprises0.2-30 wt % of glutamate equivalents, preferably in an amount of2.0-25.0 wt %, more preferably 4.0-22.0 wt %, even more preferably5.0-22.0 wt % and most preferably 8.0-21.0 wt %, in particular 10.0-20.5wt %, based on the weight of the protein fraction. Sometimes a proteinfraction containing 12.0-18 wt % of glutamate equivalents is preferred.

A relatively high weight ratio of aspartate equivalents (asp) toglutamate equivalents (glu) has a beneficial effect according to theinvention. Especially in products for young infants and babies, theweight ratio of aspartate equivalents to glutamate equivalents is ofgreat importance. Therefore, the protein fraction has a weight ratio ofasp:glu of between 0.41:1 and 5:1, preferably between 0.45:1 and 4:1,more preferably between 0.50:1 and 3:1, in particular between 0.53:1 and2:1. In another embodiment, especially for those cases where the productconsists largely of soy-based proteins, preferably more than 50 wt %,more preferably more than 60 wt %, most preferably more than 70 wt % ofthe protein fraction, an even higher asp:glu weight ratio is preferred.Then the protein fraction preferably has a weight ratio of aspartateequivalents to glutamate equivalents that is preferably between 0:58:1and 2:1, preferably in the range of 0.59:1-1.8:1, more preferably0.60:1-1.6:1, even more preferably 0.62-1.4:1 and most preferably in therange of 0.70:1-1.2:1.

Obviously better results are obtained when the extent to which thecriteria as set to the product according the invention are met becomeslarger. In particular this is true for the total amino acid compositionand the inclusion of a source of aspartate equivalents that is morerapidly available to the consumer of the product compared to the glucosefraction.

Biotin

Biotin is understood also to comprise its equivalents, i.e. allsubstances that increase blood plasma levels of D (+) biotin. Suitablesources are the acid form of D-biotin (vitamin H) and biologically andtechnologically acceptable salts or esters thereof. The amounts ofbiotin equivalents can be calculated by using the same molar amount ofthe bicyclic biotin compound. It is preferred to use food grade forms ofD-biotin.

Suitable dosages of biotin for enteral use range from 10-20000 μg perday. It is preferred that biotin dosages are 50-20000 μg per day,preferably 70-2000, more preferably 100-1000 μg/day for children above11 years and adults, and 10-500, preferably 15-250, more preferably18-150 μg for younger children. Premature infants require 6-200,preferably 8-100, more preferably 9-50 μg per daily dose. In situationswherein the disorder has a chronic nature and the enteral product isconsumed on a daily basis, like many persons suffering from diabetestype II will do in order to prevent or treat increased plasma ketonelevels, the amounts of biotin are preferably 50-1000, more preferably70-500, even more preferably 80-300 μg per day for children older than11 years and adults and 10-200, more preferably 15-150, even morepreferably 18-100 μg per day. In acute situations like acidosis, theamounts are preferably higher. For example children above 11 years ofage and adults will require 300-20000, preferably 360-2000, morepreferably 420-1000 μg per daily dose. Infants of younger age need40-500, preferably 50-250, more preferably 60-150 μg. Premature infantsshould be administered 9-200, preferably 12-100 and more preferably15-50 μg per daily dose.

The composition preferably contains biotin in an amount of 10-10000 μg,preferably 15-2000 μg, more preferably 20-1000 μg per kg, in particular50-500 μg per kg of the composition.

Biotin in the amounts according to the invention is found to decreasethe levels of ketone bodies in blood plasma, the levels of AGE productsand Maillard products in tissue, and the degree of acidosis in a shortertime, e.g. lactic acidosis but also ketonic acidosis, and to normalisethe lipid profile, in particular cholesterol plasma levels.

Vitamin B12

Vitamin B12 can be provided using suitable sources, such as syntheticCyanocobalamin, methyl cobalamin, adenosyl cobalamin and hydroxylcobalamin, for instance obtained from isolates of organs, in particularthe liver, e.g. a aqueous concentrate of lysed hepatocytes ofagricultural animals, e.g. pig, cow, chicken, with a concentrationhigher than 75 μg cobalamines per 100 ml extract.

The composition preferably contains vitamin B12 in an amount of 2.5-500μg per kg of the composition, more preferably 4-100 μg and even morepreferably 8-50 μg per kg of the composition.

When the product is to be administered to persons in the age of 3-50years and without stomach/intestinal problems or cystic fibrosis, it isuseful to ferment the product after preparation with a Lactobacillusculture, in particular a Lactobacillus acidophilus and/or L. bifidus inorder to promote the availability of vitamin B12.

Protein Fraction; Amino Acid Profile

The “protein fraction”, as used throughout the description and claims,is defined to be the sum of all proteins, peptides and amino acids inthe product, and with protein is also understood protein isolate,concentrate and/or hydrolysate. The protein fraction is effective whenit meets the following criteria:

In addition to the conditions on the amounts of aspartate and glutamateequivalents, the amount of essential amino acids in the proteinfraction, such as methionine, branched chain amino acids valine, leucineand isoleucine, and further lysine, tyrosine, phenylalanine, histidine,threonine and tryptophan, that will become available to the organism ofthe mammal after digestion of the protein fraction should providesufficient amounts to ensure anabolism and proper functioning of thebody.

In particular, it has been found that the amounts of L-methionine andL-lysine but also L-leucine are critical. Except for the case whereinthe patient suffers from tumour growth, the amount of L-methionine ispreferably 1.5-4 wt % and more preferably 1.7-3.3 wt % of the proteinfraction. Then, the sum of the amount of L-methionine and L-cysteine inthe protein fraction is preferably above 2.7 wt %, more preferably above2.9 wt % and most preferably 3.5-8 wt % of the protein fraction. In thecase that a patient suffering from net insulin resistance and/orhyperglycaemia also suffers from tumour growth, it is preferred that theprotein fraction is not supplemented with L-methionine.

Patients suffering from propionic acidemia do not tolerate high amountsof isoleucine, valine, methionine and threonine that catabolise topropionic acid in the diet. With the product of the invention the amountthat is tolerated can be increased. The total amount of these aminoacids in the protein fraction is therefore above 10, preferably 12-30,more preferably 16-26 wt % of the protein fraction. The same criterionholds for products that are used by persons that suffer frommethylmalonic acidemia.

The amount of L-lysine is preferably 5.5-15, more preferably 6.6-12 andmost preferably 7.1-11 wt % of the protein fraction. However, if it isto be administered to persons suffering from glutaric acidemia theamounts of lysine must be below 7 wt %, preferably 5.5-6.9 wt % of theprotein fraction. In that case the tryptophan levels should be below1.7, preferably 1.3-1.6 wt % of the protein fraction.

In order to avoid a large release of insulin upon administration, theconcentrations of arginine, glycine and phenylalanine in the proteinfraction must be relatively low.

The amount of arginine is preferably less than 7.9 wt %, more preferablyless than 7.8 wt %, even more preferably less than 7.0 wt % and mostpreferably less than 6.0 wt % of the protein fraction. The ratio ofL-arginine to L-lysine in the product will typically be 0.4:1-1.43:1,preferably 0.5:1-1.40:1, and especially in products to be administeredto young infants the ratio is preferably 1:1-1.40:1. The ratio ofaspartate equivalents to L-arginine in the product is preferably higherthan 1.4, more preferably 1.5-5, most preferably 1.6-3.0 to achievemaximum effect and a balanced amino acid profile.

The amount of L-glycine is preferably higher than 3.5, preferablybetween 3.6 and 4.5 wt % and more preferably less than 4.2 wt % of theprotein fraction. The weight ratio of Asp/Gly is preferably in the range2.8:1-100:1 and that of Asp/Phe in the range 2.4:1-100:1. In particularthe amount of L-serine must exceed the amount of L-glycine by at least afactor 1.5. Preferably the ratio L-serine/L-glycine is larger than 2.0:1and more preferably at least 2.3:1. This can be achieved by addingproteins that comprise a lot of L-serine relative to L-glycine and/or byadding synthetic L-serine or dipeptides which comprise L-serine.

The amount of L-phenylalanine is preferably lower than 5.6 wt % and morepreferably less than 5.3 wt % of the protein fraction. Aspartame is anunsuitable source of aspartate, also because of its extreme sweetness.

The amount of leucine in the protein fraction of the products accordingthe invention is 7.7-13 wt %. Persons that suffer from isovalericacidemia desire levels of leucine below 10 wt %, preferably below 9.0 wt%. For persons that have underdeveloped and/or imparted metabolicfunctions like young infants, pre-term infants and persons havingseverely imparted liver function, the weight ratio of aspartate relativeto leucine is preferably in the range of 0.85:1-1.5:1, more preferably0.88:1-1.4:1, even more preferably 0.9:1-1.1:1 and most preferably avalue in the range of 0.95:1-1.04:1. In order to keep the amount ofaspartate and leucine in balance it is recommended to include part ofthe amount of leucine as alpha-keto-isocaproate. This component is anexcellent counterion for components like amino acids or ornithine orbetaine in terms of effectivity and taste.

It is especially preferred to use a protein fraction satisfying thelevel of aspartate equivalents according to the invention in thepreparation of a product for treatment of metabolic disturbances,wherein the protein fraction further comprises one of: a) 7.7-19 wt % ofthe sum of all branched chain amino acids; b) 7.7-9.0 wt % leucine and3.6-4.5 wt % glycine; c) 16-26 wt % of the sum of isoleucine,methionine, valine and threonine; and d) 5.5-6.9 wt % lysine and 1.3-1.6wt % tryptophan, wherein the numbers are based on the weight of theprotein fraction.

The amount of L-histidine is preferably 2.3-4 and more preferably2.5-3.2 wt % of the protein fraction. The amount of alanine in theprotein fraction will typically be 4.8-8, preferably 5.1-7.5 and morepreferably 5.3-7.0 wt %.

Organic molecules comprising a guanidino group can be beneficiallyincluded in the product. However, it is recommended not to include freearginine or its equivalents like salts or small peptides that compriseL-arginine. Instead, low amounts of guanidino-acetate or3-guanidino-propionate can be included, e.g. in amounts below 2 g perdaily dose and preferably in amounts of 0.1-1 g per daily dose. In aliquid product 3-guanidino-propionate is an excellent source and itsconcentration will typically be 0.005-0.05 wt %. It is thereforepreferred to include no or only relatively low amounts of creatine suchthat the weight ratio of creatine/aspartate equivalents in the proteinfraction is less than 0.2:1, preferably even less than 0.1:1, morepreferably even less than 0.5:1 to avoid potentially deleterious effectsof creatine on some enzymes of the trans-sulphuration pathways. This isimportant that the product has to act on some secondary side effects ofhyperglycaemia and/or insulin resistance like some vascular disorderslike hypertension and erectile dysfunction.

As a source of methionine synthetic L-methionine, salts thereof, e.g.those with alkali metals, calcium, magnesium, zinc or organic acids likecitric acid or malic acid or amino acids like aspartic acid can be used.It is preferred to use a form that tastes better than the syntheticL-methionine. Suitable forms are acylated methionine, e.g. the N-acetylmethionine as has been described in EP 0758852 and U.S. Pat. No.1,560,000, and the methionine analogs as disclosed in U.S. Pat. No.5,430,064. A small amount of the methionine may suitably be added aszinc methioninate complex. In order to avoid that the total dose of zincexceeds 100 mg per day the amount of zinc methioninate should be below 1wt % of the protein fraction.

Embodiments

In one embodiment of the invention the nutritional or pharmaceuticalpreparation comprises a protein fraction of a first aspartate-richsource, i.e. a protein, a protein concentrate, isolate or hydrolysate oreven free aspartate equivalents, wherein the first aspartate-rich sourcecontains more than 12.0 wt %, preferably at least 12.3 wt % aspartateequivalents, and an aspartate-rich second source different form thefirst source. The second source of aspartate equivalents can be anotherprotein, preferably comprising at least 7.8 wt %, more preferably atleast 8.0 wt %, even more preferably at least 9.0 wt %, more preferablyat least 10.0 wt %, even more preferably at least 10.5 wt % aspartateequivalents. The choice of free aspartate equivalents as a first sourceis especially favoured in the case where a fast absorption of aspartatefrom the diet in the blood is required. Other preferred choices of thefirst aspartate-rich source are lactalbumine-enriched whey and potatoprotein.

It is preferred that the preparation comprises at least two proteins. Inorder to meet all the aformentioned nutritional criteria at the sametime it appears that a combination of a protein from plant origin andone of animal origin is most suitable. In addition, it appears that inthis way the resulting taste of the protein source is much better thanwhen using proteins that only consist of protein from plant origin. Theuse of the combination of a protein of plant origin and one of animalorigin also allows rapid availability of the aspartate equivalents,especially in the case where at least one of the proteins is partiallyhydrolysed. If a protein is partially hydrolysed, it is preferred thatit is the protein of plant origin, especially in case of a liquidformulation, whereas the protein of animal origin can be non-hydrolysedor only slightly hydrolysed, in order to increase solubility of theprotein and to obtain a liquid that is stable also during processing, inparticular during heating. The degree of hydrolysis is then preferably5-70%, more preferably 8-60%, most preferably 11-50%. The weight ratiobetween the protein from plant origin and the protein of animal originis preferably between 4:1 and 1:4, more preferably between 3:1 and 1:3,most preferably between 2:1 and 1:2.

Table 1 provides some comparable data which clarify the differencesbetween the protein composition according to the invention andindividual proteins known in the art.

TABLE 1 Amino acid composition of common ingredients (wt % of proteinfraction) invention soy whey^(#) EWP^(#) milk^(#) casein pea potatoinvention (optional) aspartate eq. 11.8 10.4 9.9 8.0 7.8  8.4-11^($) 2110.8-30 glutamate eq. 20.5 18.2 15.3 22.7 25.0 15.1 22.5  0.2-30 wtratio asp/glu 0.57 0.57 0.64 0.35 0.31  0.55-0.73 0.93 0.41-5 L-lysine5.6 9.2 6.5 8.8 10.2  9.3 6.4  5.5-15 L-methionine 1.6 1.9 4.3 2.7 3.3 1.5 1.5  1.5-4 L-arginine 7.8 3.0 6.2 3.6 4.0 16 5.9  1.0-7.9 L-glycine4.4 1.9 4.7 2.1 2.0  2.6 5.9  1.0-4.5 L-phenylalanine 5.5 3.2 6.9 5.15.6  6.1 4.9  3.2-5.6 L-histidine 2.5 1.6 2.3 3.0 3.2  3.4 2.0  2.3-4L-leucine 7.7 10.4 8.4 10.2 10.5  7.5 5.7  7.7-13 ^(#)whey is the bulkdesalted whey protein from cow's milk; EWP = egg white protein; milkstands for cow's milk; ^($)aspartate levels depend on the type ofspecies (see e.g. Souci, Fachmann and Kraut in Food composition andNutritional Tables, 6^(th) ed, Stuttgart, 2000), and the proteinisolation method.

When the optimal amino acid composition as disclosed in Table 1 isapplied in products for persons that suffer from an inherited metabolicdisorder, it is important that the remainder of the amino acids presentin the product complies with the specific nutritional demands of thisparticular type of patient. For example, if the product is used by aperson that suffers from Maple Syrup Urine Disease, the product shouldcomprise low amounts of branched chain amino acids e.g. less than 20 wt% of the protein, e.g. 7.7-19 wt %.

Several raw materials can be effectively used in the protein fractionaccording to the invention. Whey, soy, lupine, potato, meat, liver,fish, white bean, lima bean, lentil, pigeon pea, some other pea speciessuch as yellow Canadian pea, and black gram comprise relatively highlevels of proteins that are relatively rich in aspartate equivalentscompared to glutamate equivalents. Specific whey fractions of the milkof all mammals, in particular of cow, buffalo, horse, goat, sheep andcamel, can be used as long as they meet the above criteria. Forpractical reasons and because of its beneficial amino acid composition,whey from cow's milk is particularly suitable as starting material inmost cases, for example sweet whey that results after cheese manufactureor acid whey. The latter is a very suitable source due to the absence ofglycomacropeptide.

Raw whey from cow's milk comprises numerous proteins likebeta-lactoglobulin, immuno-globulins, lactoferrin, bovine serum albumin,alpha-lactalbumin and several others. Pure alpha-lactalbumin but alsowhey fractions that comprise more than 20% of the sum of these proteinsand preferably between 30-90 wt % and most preferably 33-70 wt % canbeneficially be used for the purpose of the invention. Very suitablewhey proteins are α-lactalbumin-enriched whey proteins having a contentof aspartate equivalents of at least 12 wt % and an asp:glu ratio of atleast 0.58, as exemplified in Table 2.

TABLE 2 Examples of the amino acid composition of two suitable wheyfractions of cow's milk for use in the products according the inventionLP^($) a-whey^(#) aspartate equivalents 12.3   13-13.5 glutamateequivalents 21.2 16.2 weight ratio asp:glu 0.58 0.80-0.83 L-lysine 10.7  9-10.1 L-methionine 2.4 1.6 L-arginine 3.0 1.8 L-glycine 2.2 2.1L-phenylalanine 3.7 3.3-3.8 L-histidine 1.6 2.4 L-leucine 11.8 12.1^(#)a-whey is a specific whey fraction isolated from cow's milk and thatis enriched in alpha lactalbumin; ^($)LP stands for a commerciallyavailable whey fraction that is enriched in alpha-lactalbumin.

Potato protein is a very suitable form of rapidly available aspartateand hydrolysis as such is not required when it is included in dryproducts. However in liquid products it should be hydrolysed in order toincrease their solubility. The same applies to proteins which arereadily digested, like meat products in non-fat nutritional products.Meat or liver proteins such as those having an aspartate equivalentcontent between 8.5 and 11 wt. % and an asp:glu ratio between 0.55 and0.9 are very suitable.

It is preferred that the preparation according to the inventioncomprises a protein from animal origin selected from meat, milk whey orliver, and a second protein from plants selected from soy, lupin, pea,in particular pigean pea, beans, in particular white bean, lima bean,lentil or black gram, and potato. It is especially preferred that theprotein fraction comprises a soy hydrolysate or concentrate, or a dairyproduct. With dairy product is understood a protein fraction thatcomprises at least 80 wt % of dairy proteins such as proteins isolatedfrom milk of cow, buffalo, camel, horse, goat, and sheep. The two mainprotein constituents of milk are whey (20 wt %) and casein (80 wt %).Such a soy protein hydrolysate or concentrate or a dairy product with anoutbalanced essential amino acid profile can be enriched with aspartateequivalents using small amounts of an aspartate-rich protein, e.g. peaprotein, potato protein or alpha-lactalbumin. The amount of such asecond protein is preferably lower than 70 wt %, more preferably lowerthan 40 wt %, even more preferably lower than 30 wt % and mostpreferably lower than 20 wt % of the protein fraction.

Several of the raw ingredients that comprise a protein fraction thatsatisfy the requirements of the invention are rich in anti-nutritionalfactors like haemaglutinins, phytic acid, tannins, flavonoids andprotease inhibitors. The amount of these components in the proteinfractions should preferably be very low, which can be achieved byapplying suitable isolation practices either alone or in combinationwith heat treatment (so called “toasting”) as is described in the art.In order to ensure that a rapidly available aspartate source is includedin the product it is important that the amount of protease inhibitorsare low, in particular when intact proteins or slightly hydrolysedproteins have been included as aspartate equivalents. The amount ofprotease inhibitors can for example be quantified as remaining TrypsinInhibitor Activity (TIA) or as concentration of Bowman-Birk inhibitorsby using methods known in the art. Typical levels are below 0.12 g,preferably below 0.06 g, more preferably below 0.02 g and mostpreferably below 0.007 g per kg of the protein fraction. In particularthe level of inhibitors of chymotrypsine should be below 0.01,preferably below 0.004 per kg of the protein fraction. The amount ofadequately treated soy protein isolates are 1-6 TIA per g of the proteinfraction.

By mixing several of the protein fractions of the ingredients asmentioned an amino acid profile can be obtained which fulfils thecriteria as set according to the invention for complete nutrition. In anembodiment of the invention mixtures of soy protein and synthetic aminoacids or soy protein with specific whey proteins, in particular wheyproteins that are enriched in alpha-lactalbumin are preferred.

It is preferred that at least one of the proteins is hydrolysed in casethat no free L-aspartic acid or salts thereof are included in theproduct, though an important part of the total protein should remainintact for taste considerations. Then typically 30-95 wt % of theprotein fraction is intact, preferably 40-92, more preferably 50-89 wt%, even more preferably at least 60 wt % and especially at least 70 wt %of the protein fraction. As described above, it is preferably that thevegetable protein source is hydrolysed rather than the protein of animalorigin, in particular because of organoleptic and product stabilityreasons, e.g. during heat treatment and/or shelf life. For example aprotein fraction that is prepared by mixing 95 wt % soy protein isolate,and 2 wt % L-aspartate and 1 wt % L-lysine and 1 wt % L-methionine wouldmeet the criteria as set.

In some embodiments it is preferred to use a large fraction of soyprotein isolate or hydrolysate. It is however preferred to use less than92 wt % of soy protein isolate, which provides about 10 wt % ofaspartate equivalents, preferably less than 90 wt % of soy proteinisolate and even more preferably even less than 85 wt % of soy proteinisolate. The protein fraction is then fortified to the required level ofaspartate equivalents using non-soy protein containing at least 12.0 wt% of aspartate equivalents or free aspartate equivalents that arerapidly digestible.

Examples of combinations of proteins satisfying the criteria of theinvention are a mixture of 83 wt % hydrolysed soy protein concentrate,15 wt % hydrolysed alpha-lactalbumin enriched whey protein (as providedby Arla) and 0.5 wt % L-methionine, 0.5 wt % L-histidine and 1 wt %L-serine, or a mixture of 40 wt % soy, 50 wt % meat protein and 10 wt %potato protein, or a mixture of 50 wt % hydrolysed soy protein isolateand 48 wt % cow's milk whey fraction, 0.5 wt % N-acetylmethionine, 0.5wt % L-histidine and 1 wt % serine.

Where it is preferred to prepare a composition on dairy basis,especially in the treatment of young infants suffering from or at riskof developing hyperglycaemia, insulin resistance or child obesity ordiabetes the amount of dairy or milk proteins is at least 50 wt % of theprotein fraction, preferably at least 60 wt %, more preferably at least70 wt % and most preferably at least 80 wt % of the protein fraction.Such a composition is to be fortified with an aspartate-rich source tomake the composition satisfy the criteria of at least an asp:glu weightratio according to the invention.

Though many components can serve as a metabolic precursor of aspartateafter digestion in the gastrointestinal tract several of thesecomponents are preferred. Intact proteins from several sources as wellas their hydrolysates are recommended. It is therefore preferred thatthe protein fraction comprises peptides, intact proteins and/orhydrolysates thereof.

Glutamate equivalents are abundantly present in the proteins as selectedto meet the requirements of the amino acids. It is however only usefulto include N-acetyl glutamine as long as the aforementioned requirementsfor the total protein composition are met and the total amount ofN-acetyl glutamine does not exceed 50 wt % of the amount of glutamateequivalents, preferably is in the range of 2-40 and more preferably 5-25weight percent of the amount of glutamate equivalents, based on theweight of the protein fraction. The latter is important to avoidhomeostasis problems with nitrogen balance. However, as this does notplay a dominant role in cases of hyper-ammonemia, the restriction on thefraction of N-acetyl glutamine does not apply when hyperammonemia isdiagnosed in a patient.

It is preferred that if the proteins are used in combination withcarbohydrates in the total diet, the amount of protein provided must beless than the amount of digestible carbohydrate. Typical amounts ofprotein in products that are meant to be used as complete nutrition willcomprise 10-30, preferably 15-25 and more preferably 18-22 energypercent, in particular about 20 energy percent.

The protein fraction will preferably comprise no or low amounts ofcaseins or its hydrolysates, because it is a poor source of aspartateequivalents and comprises too much glutamate equivalents for the purposeof the invention. The amount is preferably less than 40 wt %, morepreferably less than 25 wt % of the protein, even more preferably lessthan 10 wt % and most preferably less than 5 wt %.

In order to estimate the daily amount of the ingredients of thenutritional composition that is to be administered to achieve abeneficial effect on the levels of glucose, the protein weightpercentages throughout the text can be converted to a daily dosage usingthe following calculation, thereby assuming that the total energy supplyfor a patient is about 2000 kcal/day for a body weight of 70 kg: Atypical nutritional composition of the invention contains about 20energy percent of a protein fraction, and thus the total amount ofprotein fraction administered to a patient per day is about 400 kcal, orin weight terms, about 100 g protein fraction. Therefore, a daily dosecan be calculated on the basis on a protein consumption of 100 g per dayand, by way of example, a required asparate content of e.g. 12 wt. % ofthe protein fraction corresponds to a daily dosage of 12 g aspartate. Ifdesired, these amounts can be adapted to the actual body weight bymultiplying the required amount by B/70, wherein B is the bodyweight inkilograms. For calculating the optimal doses for an infant, an energysupply of 560 kcal and a protein content of 10 energy percent can beassumed, leading to a protein consumption of 56 kcal or 14 grams, andthis results in multiplying the required aspartate content by a factorof 0.14 (1/7). For example, a required aspartate content of 12 wt. %corresponds to a daily dosage of 12×0.14=1.68 g. If desired, theseamounts can be adapted to the body weight by multiplying by B/2, 2 kgbeing the weight of an infant used as a starting point for thesecalculations.

Carbohydrate Fraction

It is preferred to use the protein fraction in combination with at leasta carbohydrate fraction. The fraction of carbohydrates in the diet mustbe relatively slowly digested in the gastrointestinal tract of themammal compared to the protein fraction that comprises the aspartateequivalents. Best results are obtained using a product demonstrating aglycemic index below 70 and preferably below 55. This can advantageouslybe achieved using a carbohydrate fraction exhibiting a glycemic indexbelow 90, preferably between 15 and 70, more preferably between 25 and55. The glycemic index compares the immediate effect of the carbohydratefraction on plasma glucose levels compared to glucose, which is giventhe value 100. The method to determine glycemic index including valuesfor several carbohydrates is described in the art.

Suitable sources of digestible carbohydrates can be any food gradecarbohydrate extract from tubers or cereals like barley, oats, potato,corn, wheat, rye, triticale, millet, sorghum, amaranth, rice, sugarcane, sugar beet, cassaye, tapioca, etc.

The digestible carbohydrate fraction can comprise two types ofcarbohydrates: (i) the glucose equivalents, which are understood to beglucose polymers, glucose oligomers, disaccharides that comprise glucoseand glucose itself, and (ii) carbohydrates that comprise predominantlymonosaccharide units that differ from glucose. The latter category istypically difficult to digest in the gastrointestinal tract of man.However, often the monosaccharides themselves and several disaccharidesare relatively easy to absorb and to digest.

Preferably the aspartate equivalents are administered in an amountcorresponding with a weight ratio of aspartate equivalents to glucoseequivalents of 0.037:1-2:1, more preferably of 0.045:1-1.8:1, even morepreferably of 0.050:1-1.5:1 and most preferably 0.060:1-1:1. With theglucose equivalents is understood all glucose that is administered inone or more portions of the nutritional or pharmaceutical preparation,but also the equivalents that are comprised in the meal that the personconsumes within 60 minutes after administration of the aspartate-richpreparation. For the purpose of calculating the aspartate to glucoseratio, any glucose occurring in α-glucans, glucose itself, sucrose andlactose is included whether or not the glucan is readily or difficultyabsorbable or digestible.

Sources of digestible carbohydrates can be treated in such a way thatthe carbohydrates are difficult to approach by the digestive enzymes.Examples are resistant starches. The carbohydrates can also compriseglucose moieties which are attached to each other via beta-1,6- oralpha-1,1 glycosidic bonds which are difficult to hydrolyse by thenormal digestive enzymes. Examples of this type of carbohydrates havebeen described in the art, for example in WO 2004/023891, modifiedstarches and pullulan as described in WO 03/105605. Also the use ofhighly branched carbohydrates like high-amylopectin carbohydrates delaysdigestion and can suitably be included, like those starches thatcomprise more than 75 wt % amylopectin, preferably when they are lightlyhydrolysed. Suitable sources have been genetically modified or obtainedvia selection of plants like potato, tapioca, corn, cassaya or cerealslike sorghum, wheat, rye, triticale, barley, oats or millet. Othersources that can partially be included in the formula are thosemaltodextrins which comprise high amounts of polymers having more than 9monosaccharide units. By using a small degree of hydrolysis of theintact starches a suitable source of glucose is obtained. Digestion canfurther be delayed by using additives during hydrolysis of the starch,which leaves the structure of the membrane of the starch granule moreintact, like has been disclosed in U.S. Pat. No. 6,720,312.

About 40-100 wt % of the carbohydrate fraction must be formed of glucoseequivalents. Preferably this amount is 45-90, more preferably 49-80 andmost preferably 52-75 wt %. Useful glucose equivalents are for exampleglucose polymers having a chain length of more than 9 units, that forexample occur in maltodextrins DE 2-31, and some glucose syrups. Otheruseful glucose oligomers are those in which glucose occurs together withother monosaccharides like galactose, fructose, xylose, arabinose,mannose, fucose, rhamnose, sialic acid or hexuronic acids, which areincluded in an amount of 1-60 wt % of the glucose equivalents. For younginfants it is preferred to include glucose equivalents in which one offucose, rhamnose, sialic acid or hexuronic acids are included. Suitableingredients can be extracted from milk, in particular goat's milk.Examples have been given in EP0957692. For the latter group of usersthese are preferably used in an amount of 1-40 wt % of the glucoseequivalents.

Glucose polysaccharides that comprise more than 80 wt % glucose areparticularly useful for inclusion in dry products. Examples are starchtypes which demonstrate delayed digestion due to chemical or physicalmodification of the granule or the starch molecules. For the purpose ofthe invention, resistant starch can be determined by applying the methodof Englyst an Cummings, Adv. Exp. Med. Biol. 270, 205-225 (1990).Resistant starch may preferably be present at a level of 10-80,preferably 15-60, more preferably 20-40% by weight of the non-digestiblecarbohydrate (fiber) fraction.

Other examples of suitable glucose equivalents are oligosaccharides thatcomprise for more than 50 wt % glucose and that have a chain length of3-9. The amount of these oligo-glucosides should be less than 50,preferably less than 40, and most preferably less than 30% of the weightof the digestible carbohydrates. The amount of pure glucose must be lowdue to its contribution of osmotic value and its sweetness. Preferablythe amount is below 10 wt % of the carbohydrate fraction, morepreferably 1-8 wt %.

Of the category of disaccharides that comprise a glucose moiety, inparticular sucrose and lactose, it is preferred not to include sucroseat more than 5 wt % of the digestible carbohydrate fraction because ofits sweetness and contribution to the osmotic pressure of the product.Despite the fact that the latter property also applies to lactose, it ispreferred to include lactose in the product unless a clear intolerancefor lactose exists. The latter also holds for nutritional products thatcomprise a protein fraction having more than 5 wt % protein thatoriginates from plants, like soy, lupine, pea, potato, etc.

The category of monosaccharides other than glucose can also be includedin the product, though at little amounts, because they contributeheavily to osmotic value and to some extent to sweetness and may causeabdominal complaints. Examples of monosaccharides are arabinose,arabitol, mannose, ribose, galactose, rhamnose, xylulose, xylitol andfructose. The amount of hepta-carbon saccharides like sedo-heptuloseshould be less than 10 and preferably less than 5% of the weight amountof monosaccharides. The amount of the sum of all monosaccharides thatare different from glucose must be less than the amount of glucoseequivalents in the product and preferably less than 0.8 times the amountof glucose equivalents. In other words these amounts will therefore be1-40, preferably 2-30 and more preferably be 3-20 wt % of the fractionof digestible carbohydrates.

When fructose is included it is preferred to include it in relativelylimited amounts. The amount of fructose must be in the range of 0.1-20wt % of the digestible carbohydrates in order to keep the plasma levelbelow 150 and preferably below 120 microM. This is achieved preferablyby inclusion of 0.2-15 wt %, preferably 0.3-10 wt %, more preferably0.4-5 wt %, and most preferably 0.5-4 wt % of fructose, based on theweight of the carbohydrate fraction. In this way less than 2 g fructoseis consumed per meal and preferably less than 1 g. On the other hand atthe same time more than 2 g glucose units are consumed and preferablymore than 10 g per meal. The weight ratio glucose/fructose is above 2:1and preferably 5:1-100:1 and most preferably 10:1 to 50:1.

Apart from glucose and fructose also D-galactose is a preferredmonosaccharide. When the latter is included, the amount can be 1-20 andpreferably 2-10% of the weight of the monosaccharides in the products.

Digestible carbohydrates are defined to be those carbohydrates that willbe hydrolysed for more than 80% after exposure to the digestive enzymesas occur in the gastrointestinal tract and will subsequently be absorbedby the gut. The total amount of digestible carbohydrates must be 10-70energy percent, preferably 20-65, more preferably 30-60 and mostpreferably 34-55 en % of the total nutritional composition.

Using the aforementioned calculations on the conversion of the amount ofproteins in the nutrition to the daily doses thereof, thereby assuming atotal energy supply for a patient of about 2000 kcal/day, a bodyweightof 70 kg and a preferred 40 en % of digestible carbohydrates, the totalamount of digestible carbohydrates administered to a patient is about800 kcal/day, or in weight terms about 200 g digestible carbohydratesper day. It is easy for a skilled person to determine the daily dosagefor a particular patient by converting these numbers to the appropriatebody weight.

Digestion of carbohydrates can also be retarded by co-inclusion ofcomponents which decrease rate of digestion such as polyphenoliccompounds or dietary fibres. It is preferred not to include polyphenolsin the product, in order to avoid undesired interactions with proteinslike those that occur in the product or with the enzymes that act in thedigestive system. In particular the amount of flavonoids and tannins, inparticular the isoflavones, as may occur in commercially availableprotein fractions of soy or other plants, should remain below 200 mg,preferably below 100 mg and more preferably below 50 mg per daily dose.Per liter product the concentrations will therefore be less than 100 mg,preferably less than 50 and more preferably less than 25 mg polyphenolper liter product. In order to achieve this, the protein fractionsisolated from vegetable material that is rich in polyphenol content willtypically be treated for example by washing with an organic solvent likeethanol.

It is preferred that the carbohydrate fraction contains dietary fibres.Dietary fibres can be anionic polysaccharides or other poly- oroligosaccharides like for example those originating from gums likexanthan gum, Arabic gum, Konjac gum, gellan gum, tara gum and guar gum,from pectins, inulin, alginates, carragheenans, like the kappa or iotavariants, sulphated dextrans, beta-glucans especially those derived fromyeasts like Saccharomyces cerevisiae, fibers from pea, like pea hull,barley, wheat, oats or rice, or hydrolysed forms of these dietaryfibers. The fibers should have a low intrinsic viscosity in order toallow inclusion in effective amounts in a tube feeding. The viscosity ofthe final liquid form of the product needs to be 1-30 cP as measured at20° C. and at 100 per second. The use of oligosaccharides obtained byhydrolysis of the naturally occurring fibers or selection of specificisolates of the naturally occurring fibers is recommended. Effectiveamounts are typically 1-30, preferably 1.5-20 and more preferably 1.8-15g dietary fiber per daily dose for an adult. In liquid products theamounts are typically 0.05-4.0, preferably 0.075-2.5 and more preferably0.09-1.5, especially 0.1-1.0 wt % of the carbohydrate fraction. Theamount for infants can be calculated by correcting via body weight.Surprisingly it has been found that especially wheat bran orlow-methylated pectins are especially effective dietary fibres. Asdescribed above, resistant starch is an important part of the fibercomposition.

Lipid Fraction

If present, the lipid fraction should be predominantly digestible and inparticular not impart the rate of digestion and absorption of theaspartate fraction compared to the glucose equivalents.

The fatty acids within the lipid fraction predominantly have a chainlength of 18 carbon atoms or more, the so-called long chain fatty acids.In particular more than 50 wt %, preferably 60-90 wt % and morepreferably 65-80 wt % of the fatty acids are LC-fatty acids, i.e. havinga chain length of 18 or higher. The amount of unsaturated fatty acidsthat have a trans configuration is less than 0.8 wt %, preferably <0.5wt % and more preferably 0-0.3 wt % of the sum of the fatty acids. Theamount of medium chain triglycerides can be 0-20 wt % of the sum of thefatty acids and preferably 0-10 wt %. The amount of arachidonic acid isrelatively small: 0-5% and preferably 0-3% of the weight of the fattyacids. This will make the weight ratio of zinc to arachidonic acidlarger than 0.5 and preferably more than 0.8. The total amount of fattyacids in the product can be determined by extraction of the lipidfraction and determination of the fatty acids in the lipid fraction byapplying AOAC method 992.25.

Oleic acid is an important constituent in the lipid fraction. The amountis in the range of 30-60 wt % of the fatty acids. The amount of ω-3long-chain polyunsaturated fatty acids LC-PUFA's, like eicosapentaenoicacid (EPA) and docosahexaenoic acid (DHA) is relatively high. The totalamount of ω-3 LC-PUFA's is 0.5-20 wt % and preferably 1-15 wt % of thefatty acids. The sum of EPA and DHA is preferably 0.5-10 wt %, morepreferably 1-10 wt % of the fatty acids. The amount of saturated fattyacids should preferably be less than 10 wt % of the weight of the sum ofthe fatty acids.

The lipid fraction includes essential long chain fatty acids likelinoleic acid and alpha-linolenic acid as recommended by foodauthorities, in amounts of 0.8-1.5 times, preferably 1-1.2 times therequired daily dose. The amount of ω-6 LC-PUFA's, in the lipid fractionis relatively small. The amount of linoleic acid must be 5-35,preferably 6-25, more preferably 7-20 wt % of the sum of all fattyacids.

It is preferred to include the fatty acids to an important extent asphospholipids. The amount of phospholipids is 6-50, preferably 7-30 andmost preferably 8-25 wt % of the lipid fraction.

Important sources of fatty acids include structured lipids and naturaloils like marine oils like fish oil and krill extract, rice bran oil andhigh oleic vegetable oils, like olive oil and high oleic safflower oil,peanut oil and canola oil or high oleic sunflower oil extract liketrisun-80.

The total amount of lipids in complete formulae for adults andadolescents is therefore more than 30, preferably 32-60, more preferably35-50 and most preferably more than 40 energy percent of the nutritionalcomposition. In case the product is meant to be used for infants,especially premature infants, the lipids provide 30-60, preferably 31-58percent, more preferably more than 35, most preferably more than 52% ofthe total energy in the formula. This is in particular important forinfants that have underdeveloped metabolic systems like preterm babiesand for infants that are at risk for developing insulin resistance orearly obesity or diabetes, as for example becomes apparent fromprevalence of these disorders or diseases in relatives, or becomesapparent in infants with unbalanced immune systems. Examples of thelatter group of infants are those infants that have a low activity of Tcells of type 1 in relation to the activity of T cells of type 2. Thiscan be determined by measuring the amount of cytokines that are specificfor T cells type 1 (like interferon gamma) and for T cells of type 2(like interleukin-4 or 5) and comparing their weight amounts. Infantsthat demonstrate abnormal (too low) values of the weight ratioInterferon-gamma to (IL-4+IL-5), for example a ratio below 1 are definedto have an unbalanced immune system and may also lead to allergic oratopic reactions.

Product

The products according the invention can have many forms. It can be aliquid, a dry product such as a bar or a powder or a product having anintermediate moisture content such as a pudding, an ice cream or snacksof several forms. It is however preferred to use the liquid form fortube feeding and sip feeding of patients. The product can benutritionally complete or be a supplemental formula. The product can bea pharmaceutical preparation that is to be consumed simultaneously withor prior to a meal comprising glucose equivalents in order to preparethe body for the uptake of glucose from the blood. In the case theaspartate-rich nutritional or pharmaceutical preparation is to beconsumed prior to the meal, it is preferred to consume the preparationat most 60 minutes prior to a meal comprising glucose equivalents,preferably at most 45 minutes, more preferably at most 30 minutes, evenmore preferably at most 15 minutes, and most preferably at most 10minutes, especially at most 5 minutes before the meal.

A high osmolarity of the product should be avoided. The osmolarity ofthe ready to use formula is typically below 500 mOsm/1 and preferably250-400 mOsm/1. Osmolarity of the product can be measured by usingstandard methods for nutritional products known in the art. Apart fromthe rapidly available aspartate fraction the remainder of aspartateequivalents in the protein fraction can be somewhat more slowlydigestible as is the case when aspartate is present as intact proteins.For taste reasons it is much preferred to use a source of intactprotein.

Liquid complete formulae for adults and adolescents are typicallydesigned to provide 2000 kcal per day for a person weighing 70 kg, soabout 28 kcal per kg body weight per day. The volume of the formulae tobe given is therefore dependent on its energy density. When the producthas an energy density of 1.0 kcal per ml, 2 l is required to realise therequired daily doses. When the energy density is 1.25 kcal/ml about 1600ml is required per day.

Typically, the nutritional composition has an energy density of at least0.95 kcal/ml, preferably at least 1.0 kcal/ml, more preferably at least1.1 kcal/ml, and a weight ratio of aspartate equivalents to glucoseequivalents of 0.046:1-2:1, preferably at least 0.050:1, more preferablyat least 0:060:1. However, in case the nutritional composition is to beadministered to an infant, the composition preferably has an energydensity of less than 0.8 kcal/ml, more preferably less than 0.7 kcal/ml,most preferably less than 0.6 kcal/ml. With the administration of thecomposition a weight ratio of aspartate equivalents to glucoseequivalents of 0.037:1-2:1, preferably at least 0.040:1, more preferablyat least 0:045:1 and most preferably at least 0:050:1 is accomplished.Herein, the weight ratio of asp:glucose is based on the amounts ofaspartate and glucose equivalents that are present in the composition aswell as those given in a meal within 60 minutes after administration ofthe composition, the numbers being based on the total weight of proteinsand carbohydrates, respectively.

For infants the amount of energy that is provided per day is about 540kcal for an infant that weighs 3 kg, so about 180 kcal/kg body weightper day. This amount of energy rapidly decreases with increasingbodyweight to an amount of about 60 kcal/kg body weight per day afterseveral months of age. When the product is a supplement that supportscomplete nutrition, and prevents hypo- and hyperglycaemia and/or insulinresistance, the amount of energy that will be provided per day will bein the range of 100-800, preferably 180-600 and more preferably 190-560kcal. When the product is used as a nutritional or pharmaceuticalcomposition in combination with existing meals, the amount of energyprovided will be 10-200 kcal per dose, preferably 15-160 kcal and morepreferably 20-140 kcal per dose. This also applies for the case wherethe product is applied simultaneously with or prior to a meal comprisingglucose equivalents.

Infant formulae are defined to be nutritional products intended forcomplete nutrition of babies or infants from birth to an age of 24months after birth and that comprise 6-12.5 en % of a protein fraction,38-50 en % digestible carbohydrates, 40-52 en % of a lipid fraction andall minerals, trace elements and vitamins according to the officialrecommendations in an amount of 0.8-1.2 times the recommended dailyintakes per daily dose and have an energy density of 55-76 kcal permilliliter.

The nutritional composition can comprise at least two separate portions,wherein one portion comprises a protein-rich fraction and a relativelycarbohydrate- and fat-poor fraction and another portion comprisesrelatively a lot of glucose equivalents and less proteins on a weightbasis than the first portion, and which portions are administeredsequentially and wherein the portion comprising the protein-richfraction is administered not earlier than 60 minutes beforeadministration of a carbohydrate-rich fraction. Preferably the timebetween the administration of the protein-rich fraction and thecarbohydrate-rich fraction is less than 45 minutes, preferably less than30 minutes, more preferably less than 15 minutes, even more preferablyless than 10 minutes and most preferably less than 5 minutes, whereinthe portion comprising the protein-rich fraction is given first. The twoportions together satisfy the aforementioned criteria of the nutritionalproduct of the invention.

In the case of sequential administration, it is preferred that theprotein level in the first portion is typically larger than the amountof digestible carbohydrates in terms of energy. Typically the proteinlevel is 40-80 en % in the first portion, whereas the carbohydratefraction in the first portion is lower than 60 en %, preferably lowerthan 50 en %, most preferably lower than 40 en % based on the totalenergy content of the first portion. In liquid formulae this firstportion will comprise 8-10 wt % protein fraction and the amount ofdigestible carbohydrates 5-15 wt %, preferably 6-12 wt %, based on thetotal weight of the first portion, including the liquid. In relativelydry form the first portion can take the form of a snack or a bar. It ispreferred to include dietary fibre in an amount of 3-30 wt % of the drymass of the first portion.

The second portion can be any regular food product that comprises aglucose source. Typically this second portion will comprise 10-32,preferably 14-30 and more preferably 18-22 en % protein, based on theenergy content of the second portion. The carbohydrates contribute25-70, preferably 30-60, more preferably 34-56, most preferably 38-54 en% of the second portion. The lipid fraction originates for 80-100% fromthe second portion in the diet, which amounts typically to 20-130 gramlipids.

The contribution of proteins, carbohydrates and lipids to the energycontent of a product is calculated by using the methods known in theart, using the factors 4 kcal per gram protein equivalent or digestiblecarbohydrate equivalent and using the factor 9 kcal per gram lipidswhich include the phospholipids.

It is preferred that the enteral composition provides more than 1800kcal per day, more preferably 1900-2500 kcal/day, preferably about 2000kcal/day for adults and adolescents. If the composition is used foradministration to premature infants, the composition provides more than225, preferably 300-1000 kcal/day.

Minerals etc.

The nutritional composition according to the invention optionallycomprises other components than the aforementioned protein, digestiblecarbohydrate and lipid fractions. Below several components arementioned, including preferred ingredients and doses.

In those embodiments wherein arginine levels in the protein fraction arerelatively low, e.g. below 4.0 wt % and certainly below 3.0 wt % of theprotein fraction, it is advised to include L-ornithine and/orL-citrulline in the product. It is preferred that the amount of arginineplus ornithine and any citrulline is at least 3.0, especially at least4.0 wt % of the protein fraction. It is preferred to use L-ornithine orits equivalents in a ratio L-ornithine/citrulline>1 and preferably >5.The L-isomers are preferred. Recommended amounts are 0.3-5 wt % andpreferably 0.5-4 wt %, based on the weight of the protein fraction. Theweight ratio of L-ornithine+L-citrulline to L-arginine is in the range0.07:1-2:1 and preferably 0.12:1-1.2:1. The amount of L-ornithine toL-arginine in the product comprising intact proteins and/or hydrolysedforms thereof will therefore be in the range of 0.11-1.1 and preferably0.2-0.9. L-ornithine can also be included as an extract from rawingredients like meat or liver. Suitable forms are also salts, inparticular those with organic acids like amino acids for example theaspartate salt, or organic acids like malic acid or citric acid orα-keto-isocaproate (or 2-oxo-isocaproate).

By inclusion of additional L-ornithine and/or L-citrulline or theirequivalents, in particular in combination with the supplementedmethionine equivalents, endogenous polyamine biosynthesis rate isensured. Inclusion of additional ornithine or its equivalents into theformula supports renal function in persons suffering from hyperglycaemiaor insulin resistance. In order to further increase these effects it isimportant to include carbonates or bicarbonates in the product. Suitableforms are the salts with metals like sodium, potassium, lithium,magnesium, zinc, iron, copper and calcium. The use of cupric carbonate,calcium carbonate and bicarbonates of sodium, magnesium and potassium isrecommended. The pH of the formula must be in the range 6.3-7.1 andpreferably in the range 6.4-6.8. The amount of carbonates andbicarbonates, including the counter ion must be in the range of 0.8-10,preferably 1.0-6 g and more preferably 1.2-5 g per 100 g dry mass of theformula.

In patients suffering from insulin resistance or increased levels inblood of glucose, resulting in renal complications or a disorder inkidney function, the levels of biotin must be increased to a levelbetween 40 and 4000 μg/100 ml. Magnesium should be included in aconcentration of 4-20 mg/100 ml liquid product according the invention.The protein levels in this embodiment of the invention must be between10 and 22 energy percent of the composition.

The nutritional products according the invention demonstrate essentiallyno hormone activity when consumed per orally. Hormone-type componentsselected from glucagons and steroidal compounds are therefore present inamounts less than 10 mg glucagon per liter product. Levels of steroidsare typically below 0.1 ppm and preferably non-detectable.

Where the protein fraction exhibits a weight ratio of serine to glycineof less than 2.3:1, a component selected from the group choline,betaine, dimethylglycine and sarcosine must be included in order tosupport effectiveness in the treatment of hyperglycaemia and during netinsulin resistance, in particular in those patients that suffer alsofrom malnourishment and inflammation. The daily dose of these componentsshould be more than 0.5 g and preferably more than 0.8 g. In a liquidproduct according the invention the concentration becomes therefore morethan 0.025 wt % or preferably 0.032-2, more preferably 0.04-0.4 wt % andmost preferably 0.06-0.25 wt. %. In dry products the amount willtypically be 0.04-3 wt %. Effectiveness can be established viameasurement of circulating markers of inflammation like blood levels ofC-reactive protein or of several cytokines.

It is important that the product will not demonstrate extensivemaillardation i.e. browning, during manufacture, and in particularduring sterilisation. This is achieved by preventing inclusion ofcomponents like carnosine next to reducing sugars in liquid products.The weight ratio of L-lysine to carnosine in the product is thereforetypically larger than 5:1 and preferably larger than 10:1.

Zinc is an essential mineral for persons that suffer from hyperglycaemiaand/or insulin resistance. The amount of zinc is typically more than 14mg, preferably more than 18-40 mg, more preferably 20-35 mg and mostpreferably 22-30 mg per daily dose. It is important to keep the amountof copper relatively low, e.g. in a weight ratio zinc to copper of7-16:1, and preferably 8-15:1 and most preferably 9-13:1. Despite therelatively high concentration of zinc in the product, the weight ratioof zinc to L-histidine in the product is, due to the relatively highamount of L-histidine, preferably in the range 0.002:1-0.2:1.

Calcium can advanaeously be included in an amount of more than 40 mg,preferably 50-200 and more preferably 60-120 mg per 100 ml.

Magnesium can be included in liquid formulae in a dose of 20-60,preferably 25-40 and more preferably 28-35 mg per 100 ml liquid product.Magnesium triphosphate, carbonate and bicarbonate are suitable sourcesof magnesium for use in liquid formulae.

Sodium levels are typically less than 100, preferably 50-80 and morepreferably 55-74 mg per 100 ml liquid product according the invention.The weight ratio of sodium to potassium will be typically 0.3-0.66,preferably 0.4-0.64 and more preferably 0.45-0.62.

Chromium or vanadium should be included in an amount of 1-50 μg per 100ml liquid product according the invention.

It is important that in complete diets all vitamins, minerals and traceelements are included in sufficient amounts to meet nutritionalrequirements as for example set by the Food and Drug Administration andat the same time not exceed these recommendations in order to avoidoverdoses during longer term and frequent use, except where indicated inthe description.

It is preferred to include vitamin B6 in the nutritional composition ofthe invention. The levels are preferably selected to be at least twotimes the recommended daily amounts to further improve the effect of theproduct on the treatment of hyperglycaemia and/or insulin resistance.

Pyridoxine, pyridoxamine or pyridoxal or their salts, phosphorylated,glycosylated or other derivatives, either prepared synthetically orisolated from natural sources can be used as suitable sources of vitaminB6 and in particular pyridoxine. It is preferred to include 3.2-100 andpreferably 3.5-30 mg of vitamin B6 or a source thereof per daily dosefor an adult. The weight amount of vitamin B6 in the formula will beless than the weight amount of aspartate equivalents or magnesium in theproduct. Typically the amount of vitamin B6 is less than 0.01 times theamount of aspartate equivalents in the product and less than 0.1 timesthe amount of magnesium. For a complete infant formula, the amount ofvitamin B6 is preferably more than 75 μg per 100 kcal, especially 80-120μg/100 kcal.

It is further recommended to include relatively high levels ofpantothenic acid and lipoic acid. Pantothenic acid should be included inan amount of 12-300, preferably 14-100 and most preferably 18-40 mg perdaily dose as acid or its salts or pantethine or pantothenol for anadult. Per 100 ml liquid product according the invention the amounts aretherefore 0.6-15, preferably 0.7-5 and most preferably 0.9-2 mg. For acomplete infant formula, the preferred amount of pantothenic acid ismore than 480 μg, especially 500 μg-2.0 mg. Lipoic acid can be includedin an amount of 5-500, preferably 10-300, and most preferably 20-200 mgper daily dose, in forms that are known in the art like the free salt,salts thereof or better tasting derivatives. Per 100 ml of a liquidproduct according to the invention the amounts are therefore 0.25-25,preferably 0.5-15 and most preferably 1-10 mg lipoic acid.

Folic acid, salts thereof or methylated derivatives thereof arepreferably included in an amount of 300-3000, preferably 350-2000 morepreferably 400-1500 and most preferably 500-1200 microgram per dailydose for an adult. Per 100 ml liquid product according the invention theconcentration of folic acid is therefore 15, preferably 17.5-100, morepreferably 20-75 and most preferably 25-60 microgram. In a completeinfant formula, the preferred amount of folic acid is above 18 μg per100 kcal, especially 19-40 mg per 100 ml. Hiasgftugf. nhj lkf

When the present nutritional product is intended to be administered to ayoung infant who may suffer from underdeveloped metabolic systems, it ispreferred to include also limonene. This compound can be given as pure(R)-(+)-limonene as prepared synthetically or as isolated from fruitslike citrus fruits. This isolation is preferably applied via steamdistillation. The concentration should be in the range 1-1000 mg per 100g dry matter of the product.

The composition of the invention can be consumed in combination withinsulin. The composition is found to beneficially reduce the requireddosage of insulin, therewith also reducing the risk of insulinresistance.

Treatment

The product is suitable for the prevention and/or treatment of metabolicdisorders, associated with elevated concentrations of ketone bodies,lactate and/or other organic acids in blood, and/or insufficient pHhomeostasis, preferably those associated with elevate concentrations ofketone bodies and lactate, and to prevent and or treat secondarydisorders associated with these metabolic disorders.

The metabolic disorders in the context of the invention comprisemetabolic acidoses and long-term advanced glycation product (AGE)formation and/or Maillard product formation in tissue, in which caseincreased levels of glycosylated products, in particular of HbA1c, areobserved in blood and/or tissue. The presence of high levels of AGE andMaillard products in the body may contribute to the occurance ofdementia syndromes, retinopathies and transient ischaemic accidents.

Metabolic acidoses in the context of the invention are caused byincreased levels of ketone bodies and/or lactate which are short-lastingand occur in acute situations, and typically comprise hyperketonaemia,(hyper)ketosis, ketoacidosis, (hyper)ketonuria, hyperlactacidemia,organic acidurias such as lactic acidaemia and lactic acidosis. Abnormalconcentrations of ketone bodies, ammonia, lactate or other organicacids, pyruvate, glucose, a low blood pH and/or partial carbon dioxidepressure (PCO₂) are important indicators thereof. The treatment ofmetabolic acidoses is in particular of importance for neonates and inparticular preterm babies, and patients during and after surgery.

Mammals in need of the product of the invention typically suffer fromdisorders described in the introduction, in particular with secondarydiseases or disorders from the group of cardiovascular diseases, inparticular atherosclerosis and microvascular problems, cerebro-vascularproblems in particular Transient Ischemic Accidents and CerebroVascularAccidents, renal diseases, obesitas, childhood obesitas, imparted eyesight, high blood pressure and loss of tissue or organ function,imparted immune function, dysfunction of the sexual organs, inparticular imparted libido, catabolism especially after trauma, surgeryor during severe phases in diseases like cancer, infection,gangrene-type problems with limbs, acquired immune distress syndrome,metabolic syndrome, diabetes, increased HbA1C levels, chronicinflammation, chronic obstructive pulmonary disease and liver diseases.

A mammal in need of the nutritional or pharmaceutical composition of theinvention may be suffering from hyperglycaemia after fasting orpostprandially, insulin resistance, diabetes, but also from inheritedmetabolism errors such as maple syrup disease, inherited disorders inglycogen synthesis rate, propionic acidemia, isovaleric acidemia,methylmalonic acidemia, oxoacid coenzym A thiolase deficiency ordeficiencies in the activity of other thiolases, or may be a personhaving an underdeveloped metabolic system like infants of younggestational age.

The product is especially suitable for the female gender, where insulinresistance plays an important role.

The effect of the product can be determined by measuring the levels ofglucose in blood after consumption of the nutritional product accordingthe invention. Consumption of the protein fraction comprising therelatively high amount of rapidly available aspartate will decreasepostprandial or post-surgery glucose levels as observed afterconsumption of a glucose source. This is particularly the case when theaspartate is more rapidly available to the organism than the glucosesource. In particular the slow release system of the carbohydratefraction as disclosed ensures a more slow availability of the dieteticglucose compared to the aspartate. Another way of achieving this effectis through sequential administration of separate protein and digestiblecarbohydrate fractions, wherein the protein fraction is administeredprior to the carbohydrate fraction. Clearance rate of glucose andinsulin (t½) can be monitored as well as the effect on steady stateglucose and insulin levels. The effectiveness of the dietetic regimenbecomes for example clear from the number of times a hypoglycaemic state(blood glucose below 50 mg/100 ml) occurs, in combination with thenumber of times a hyperglycaemic state occurs.

The effectiveness of the inclusion of the amino acids as specified inthe description can be determined by measuring loss in lean body mass inpersons that suffer from hyperglycaemia and insulin resistance and inparticular in malnourished diabetic patients.

The effect of the inclusion of the vitamins as specified in thedescription can be determined by measuring rate of lipolysis, insulinresistance and lean body mass in obese persons that suffer fromhyperglycaemia and/or insulin resistance.

Risk for complications can be monitored by measuring the decrease inblood levels of HbA1c and/or C-reactive protein. Incidence of typicalcomplications like cardiovascular problems can be tabulated. In personsthat suffer from hyperglycemia, insulin resistance or increased bloodlevels of ketobodies and additional renal problems blood ammonia levelsshould be measured.

In persons suffering from increased insulin resistance due to increasedrelease of stress hormones plasma glucose levels and changes in leanbody mass after experiencing of e.g. the trauma or severe surgery shouldbe measured, including the morbidity and mortality.

The effect of the product on the development of acidosis can bemonitored by measuring for example blood levels of gases, electrolyteslike bicarbonate and sodium, pyruvate, lactate, glucose and pH.

It is important to extend the measurements to a period of 2 days orlonger in order to allow adaptation of the organism to the dieteticprotein composition. The effect thereafter can however be immediate oron longer term, dependant on the patient.

EXAMPLES Example 1

A drink for enteral use in the prevention or treatment of metabolicacidosis comprising 1.0 kcal/ml, 17 en % protein fraction consisting of50 wt % alpha-lactalbumin and 50 wt % soy protein isolate, 45 en %carbohydrate fraction and 38 en % lipid fraction. It further comprises30 μg biotin and 10 μg hydroxycobalamine per 100 ml product, and 16 mgMagnesium, 4 mg zink and 80 mg calcium per 100 ml product. 200 ml of thedrink is administered 2-5 times a day.

Example 2

Drink for enteral use in prevention of increased lactate levels intissue, providing 1.0 kcal/ml, 22 en % protein fraction comprising 79 wt% alpha-lactalbumin enriched whey, 1 wt % aspartate dipeptide (arginineor serine) and 20 wt % soy protein isolate, 55 en % carbohydrates, 23 en% lipids, a vitamin fraction providing per 100 ml 50 μg biotin and 10 μgvitamin B12 and a mineral/trace element fraction providing calcium,magnesium and zinc in a concentration within the range according to thedescription. The drink must be consumed in a volume of 0.5-2 litre perday.

Example 3

Drink for enteral use for treatment or prevention of hyperketonemia,providing 1.0 kcal/ml and 18 en % protein, comprising 60 wt %alpha-lactalbumin enriched whey and 40 wt % soy protein, 2 wt %aspartate-enriched potato protein, 35 en % lipids and 47 en %carbohydrate fraction and a vitamin fraction that provides per 100 ml 20μg biotin and 5 μg Cyanocobalamine. The drink must be consumed in avolume of 200 ml-2 l.

Example 4

Drink for enteral use by obese persons, providing 1.0 kcal/ml, 20 en %protein comprising 63 wt % soy protein, 15 wt % aspartate enrichedpotato protein and 20 wt % whey protein, 2 wt % amino acids (L-lysine,L-methionine, L-serine), 35 en % lipids, 45 en % carbohydrates and avitamin fraction that provides per 100 ml 20 μg biotin equivalents and10 μg vitamin B12.

1. An enteral nutritional composition comprising a protein fraction anda carbohydrate fraction providing 15-22 en % and 25-50 en %,respectively, wherein the protein fraction comprises at least 10.8 wt %of aspartate equivalents, based on the total weight of the proteinfraction, and wherein the composition further comprises at least one ofvitamin B12 and biotin.
 2. The nutritional composition according toclaim 1, comprising vitamin B12 in an amount of 2.5-500 μg per kg of thecomposition.
 3. The nutritional composition according to claim 1,comprising biotin in an amount of 10-10000 μg per kg of the composition.4. The nutritional composition according to claim 1, wherein the proteinfraction comprises an aspartate source comprising at least 12.0 wt %aspartate equivalents.
 5. The nutritional composition according to claim4, wherein the aspartate source comprises free aspartate equivalents inan amount of 0.2-9 wt % of the protein fraction.
 6. The nutritionalcomposition according to claim 4, wherein the aspartate source is aprotein, or isolate, concentrate of hydrolysate thereof, which ispresent in an amount of 5-100 wt % of the protein fraction.
 7. Thenutritional composition according to claim 5, wherein the protein is alactalbumin-enriched whey or a potato protein.
 8. The nutritionalcomposition according to claim 1, wherein the protein fraction furthercomprises glutamate equivalents in a weight ratio of aspartateequivalents to glutamate equivalents (asp:glu) between 0.41:1 and 5:1.9. The nutritional composition according to claim 1, the compositionbeing in liquid form.
 10. The composition according to claim 1, furthercomprising a carbohydrate fraction comprising 40-100 wt % glucoseequivalents.
 11. The composition according to claim 1, wherein thecomposition has a protein fraction comprising a protein fractioncomprising 12.0-40 wt % of asparate equivalents, based on the totalweight of the protein fraction.
 12. The composition according to claim1, wherein the composition has an energy density of less than 0.8kcal/ml and comprises glucose equivalents, wherein the weight ratio ofaspartate equivalents to glucose equivalents is in the range of0.037:1-2:1.
 13. The composition according to claim 1, wherein thecomposition has an energy density of at least 0.95 kcal/ml and comprisesglucose equivalents, wherein the weight ratio of aspartate equivalentsto glucose equivalents is in the range of 0.046:1-2:1.
 14. A method forprevention or treatment of diabetes, comprising administering to amammal in need thereof a composition comprising: (i) a protein fractioncomprising at least 10.8 wt % aspartate equivalents, based on the totalweight of the protein fraction; (ii) vitamin B12, biotin, or both. 15.The method according to claim 14, wherein the diabetes is type I, typeII or gestational diabetes.
 16. A method for prevention and/or treatmentof metabolic disorders associated with elevated concentrations of ketonebodies, lactate and/or organic acids and/or insufficient pH homeostasisin a mammal's blood, comprising administering to a mammal in needthereof a composition comprising: (i) a protein fraction comprising atleast 10.8 wt % aspartate equivalents, based on the total weight of theprotein fraction; (ii) vitamin B12, biotin, or both.
 17. The methodaccording to claim 16, wherein the protein fraction provides 15-22 en %.18. The method according to claim 16, wherein the protein fractioncomprises lactalbumin-enriched whey or a potato protein.